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目的:探讨反义胸腺素α原(Prothymosin alpha,ProTα)基因转染对胃癌细胞生长的影响,为以ProTα为靶向的胃癌基因治疗开辟新的途径。方法:人工合成ProTα反义寡核苷酸(ProTα-AS-ODN),以阳离子聚合物转染法转染体外培养的人胃癌细胞BGC-823,以正义寡核苷酸和未转染组为对照,应用RT-PCR及免疫细胞化学(immunocytochemistry,ICC)法分析ProTα基因及其蛋白的表达,MTT法分析细胞生长抑制作用,Hoechst33258检测细胞凋亡。结果:RT-PCR、ICC显示ProTα-AS-ODN转染组ProTα mRNA及其蛋白的表达显著减少,MTT检测表明ProTα-AS-ODN转染组活细胞数较其它各组均低(P<0.05),Hoechst33258则显示其凋亡细胞数较其它各组明显增高(P<0.01)。结论:ProTα-AS-ODN成功转染人胃癌细胞BGC-823,封闭了ProTαmRNA的翻译,使已转染的细胞ProTα蛋白的表达减少,且可抑制细胞的增殖并诱导细胞发生凋亡。
Objective: To investigate the effect of Prothymosin alpha (ProTα) gene transfection on the growth of gastric cancer cells and to explore a new approach for the gene therapy of gastric cancer targeting ProTα. Methods: Human ProTα antisense oligodeoxynucleotide (ProTα-AS-ODN) was synthesized and transfected into BGC-823 human gastric cancer cells by cationic polymer-mediated transfection. The expression of ProTα gene and its protein was analyzed by RT-PCR and immunocytochemistry (ICC). Cell growth inhibition was detected by MTT assay and apoptosis was detected by Hoechst33258. Results: RT-PCR and ICC showed that the expression of ProTα mRNA and protein in ProTα-AS-ODN transfected group was significantly decreased. The number of viable cells in ProTα-AS-ODN transfection group was lower than that in other groups (P <0.05 ), Hoechst33258 showed that the number of apoptotic cells was significantly higher than other groups (P <0.01). Conclusion: ProTα-AS-ODN was successfully transfected into BGC-823 human gastric cancer cells, which blocked the translation of ProTα mRNA and decreased the expression of ProTα protein in transfected cells, and inhibited the proliferation of cells and induced cell apoptosis.